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Lehigh University

14 ARTICLES PUBLISHED IN JoVE

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Biology

PDMS Device Fabrication and Surface Modification
Kenneth Kotz 1, Xuanhong Cheng 1, Mehmet Toner 1
1Havard Medical School, MGH - Massachusetts General Hospital

PDMS Device Fabrication and Surface Modification

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Biology

Cell Capture Using a Microfluidic Device
Kenneth Kotz 1, Xuanhong Cheng 1, Mehmet Toner 1
1Havard Medical School, MGH - Massachusetts General Hospital

Cell Capture Using a Microfluidic Device

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Biology

An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila
Daniel T. Babcock 1, Barry Ganetzky 1
1Department of Genetics, University of Wisconsin-Madison

Here we describe a method for rapid and accurate measurement of flight performance in Drosophila, enabling high-throughput screening.

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Bioengineering

Formation of Biomembrane Microarrays with a Squeegee-based Assembly Method
Nathan J. Wittenberg 1, Timothy W. Johnson 1, Luke R. Jordan 2, Xiaohua Xu 3, Arthur E. Warrington 3, Moses Rodriguez 3,4, Sang-Hyun Oh 1,2
1Department of Electrical and Computer Engineering, University of Minnesota, 2Department of Biomedical Engineering, University of Minnesota, 3Department of Neurology, Mayo Clinic College of Medicine, 4Department of Immunology, Mayo Clinic College of Medicine

Supported lipid bilayers and natural membrane particles are convenient systems that can approximate the properties of cell membranes and be incorporated in a variety of analytical strategies. Here we demonstrate a method for preparing microarrays composed of supported lipid bilayer-coated SiO2 beads, phospholipid vesicles or natural membrane particles.

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Chemistry

Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
Mary J. Sabulski 1, Jonathan M. Fura 1, Marcos M. Pires 1
1Department of Chemistry, Lehigh University

PAD4 is an enzyme responsible for the conversion of peptidyl-arginine to peptidyl-citrulline. Dysregulation of PAD4 has been implicated in a number of human diseases. A facile and high-throughput compatible fluorescence based PAD4 assay is described.

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Biology

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)
Jing Men 1,2, Jason Jerwick 2,3, Penghe Wu 1,2, Mingming Chen 3,4, Aneesh Alex 2,3, Yutao Ma 4, Rudolph E. Tanzi 5, Airong Li 5, Chao Zhou 1,2,3
1Bioengineering Program, Lehigh University, 2Center for Photonics and Nanoelectronics, Lehigh University, 3Department of Electrical and Computer Engineering, Lehigh University, 4State Key Laboratory of Software Engineering, Wuhan University, 5Genetics and Aging Research Unit, Department of Neurology, Massachusetts General Hospital and Harvard Medical School

Here, the experimental protocols are described for preparing Drosophila at different developmental stages and performing longitudinal optical imaging of Drosophila heartbeats using a custom optical coherence microscopy (OCM) system. The cardiac morphological and dynamical changes can be quantitatively characterized by analyzing the heart structural and functional parameters from OCM images.

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Genetics

Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
Sara Lynn N. Farwell 1, Joshua B. Slee 2, Yaqiu Li 1, Linda J. Lowe-Krentz 1
1Department of Biological Sciences, Lehigh University, 2Department of Natural Science, DeSales University

A construct encoding TMEM184A with a GFP tag at the carboxy-terminus designed for eukaryotic expression, was employed in assays designed to confirm the identification of TMEM184A as a heparin receptor in vascular cells.

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JoVE Core

Combining Microfluidics and Microrheology to Determine Rheological Properties of Soft Matter during Repeated Phase Transitions
Matthew D. Wehrman 1, Melissa J. Milstrey 1, Seth Lindberg 2, Kelly M. Schultz 1
1Department of Chemical and Biomolecular Engineering, Lehigh University, 2Process and Engineering Development, Procter & Gamble Co.

We demonstrate the fabrication and use of a microfluidic device that enables multiple particle tracking microrheology measurements to study the rheological effects of repeated phase transitions on soft matter.

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Cancer Research

Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography
Yongyang Huang 1, Jinyun Zou 1, Mudabbir Badar 1, Junchao Liu 1, Wentao Shi 5, Shunqiang Wang 2, Qiongyu Guo 3, Xiaofang Wang 1, Sarah Kessel 4, Leo Li-Ying Chan 4, Peter Li 4, Yaling Liu 2,5, Jean Qiu 4, Chao Zhou 1,5,6
1Department of Electrical and Computer Engineering, Lehigh University, 2Department of Mechanical Engineering, Lehigh University, 3Department of Biomedical Engineering, Southern University of Science and Technology, 4Department of Technology R&D, Nexcelom Bioscience LLC, 5Department of Bioengineering, Lehigh University, 6Center for Photonics and Nanoelectronics, Lehigh University

Optical coherence tomography (OCT), a three-dimensional imaging technology, was used to monitor and characterize the growth kinetics of multicellular tumor spheroids. Precise volumetric quantification of tumor spheroids using a voxel counting approach, and label-free dead tissue detection in the spheroids based on intrinsic optical attenuation contrast, were demonstrated.

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Bioengineering

Characterizing Single-Molecule Conformational Changes Under Shear Flow with Fluorescence Microscopy
Avani V. Pisapati 1, Yi Wang *2, Megan E. Blauch *3, Nathan J. Wittenberg 3, Xuanhong Cheng 1,2, X. Frank Zhang 1,4
1Department of Bioengineering, Lehigh University, 2Department of Materials Science and Engineering, Lehigh University, 3Department of Chemistry, Lehigh University, 4Department of Mechanical Engineering and Mechanics, Lehigh University

We present a protocol for immobilizing single macromolecules in microfluidic devices and quantifying changes in their conformations under shear flow. This protocol is useful for characterizing the biomechanical and functional properties of biomolecules such as proteins and DNA in a flow environment.

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Neuroscience

Visualizing Synaptic Degeneration in Adult Drosophila in Association with Neurodegeneration
Jessica M. Sidisky 1, Daniel T. Babcock 1
1Department of Biological Sciences, Lehigh University

The goal of this procedure is to dissect the dorsal longitudinal muscle (DLM) tissue to assess the structural integrity of DLM neuromuscular junctions (NMJs) in neurodegenerative disease models using Drosophila melanogaster.

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Bioengineering

Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
Shannon M. Collins 1, Justin B. Nice 1, En Hyung Chang 1, Angela C. Brown 1
1Department of Chemical and Biomolecular Engineering, Lehigh University

Bacterial vesicles play important roles in pathogenesis and have promising biotechnological applications. The heterogeneity of vesicles complicates analysis and use; therefore, a simple, reproducible method to separate varying sizes of vesicles is necessary. Here, we demonstrate the use of size exclusion chromatography to separate heterogeneous vesicles produced by Aggregatibacter actinomycetemcomitans.

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Bioengineering

Developing Drosophila melanogaster Models for Imaging and Optogenetic Control of Cardiac Function
Elena Gracheva 1, Fei Wang 1, Abigail Matt 1, Hongwu Liang 1, Matthew Fishman 1,2, Chao Zhou 1
1Department of Biomedical Engineering, Washington University in St. Louis, 2Department of Computer Science and Engineering, Washington University in St. Louis

The present protocol describes the generation of Drosophila melanogaster expressing eNpHR2.0 or ReaChR opsins in the heart for OCT imaging and optogenetic heart pacing. Detailed instructions for Drosophila OCT imaging and heart beating modulation, including the simulation of restorable heart arrest, bradycardia, and tachycardia in live animals at different developmental stages, are reported.

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Chemistry

An Introduction to Processing, Fitting, and Interpreting Transient Absorption Data
Robert Hamburger 1, Christopher Rumble 2, Elizabeth R. Young 1
1Department of Chemistry, Lehigh University, 2Department of Chemistry, Pennsylvania State University - Altoona College

This protocol is a beginner's entryway into processing, fitting, and interpreting transient absorption spectra. The focus of this protocol is the preparation of datasets, and fitting using both single wavelength kinetics and global lifetime analysis. Challenges associated with transient absorption data and its fitting are discussed.

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